of Microbial Community Structure in Estuarine Environments using Flow Cytometry
Brian Binder, Dept of Marine Sciences, Univ of Georgia, Athens, GA, USA
Georgia Sea Grant College Program (R/AT-2)
3/1/97 - 6/1/98 (complete)
To develop methods using flow cytometry and DNA-specific fluorescent stains
to detect and enumerate different bacterial groups based on their DNA base composition.
- Sample preservation
and staining protocols were developed and found to be satisfactory for all
bacterial strains examined. These strains included: Altermonas macleodii
(%G+C = 46), Escherichia coli [used as a control](%G+C = 50%), Delaya
venusta (%G+C = 54%), Pseudomonas doudoroffii (%G+C = 60%), Sagittula
stellata (65%), and Delaya pacifica (%G+C = 68%). The DNA stains
Hoechst 33342 (HO) and Chromomycin A3 (CA3) were used as AT- and GC-specific
- The strong correlation
between the measured CA3:HO ratios and %G+C suggests that this analytical
approach holds considerable promise.